Molecular & Genetic Determinants of Outcome in Breast Ca

描述(申请人提供)：乳腺癌仍然是一个主要的公共卫生问题，美国每年有超过19万例新病例。这些病例中约有三分之一是女性，其疾病仅限于乳房和腋窝淋巴结。在这些患者中，积极的多模式治疗可以在略高于50%的病例中治愈。然而，目前的预后标记物无法区分那些将通过治疗治愈的人，那些没有积极治疗就被治愈的人，以及那些无论治疗如何都注定会复发的人。为了提高生存率和适当的靶向治疗，显然需要更好的标记物和对复发生物学的更好理解。这项拟议的研究旨在通过研究宿主遗传特征对治疗结果的作用，以及这些宿主因素与肿瘤分子特征之间的联系，来扩大我们对乳腺癌复发和预后的了解。这项研究中感兴趣的宿主遗传因子是白介素6(IL-6)，它是一种参与宿主对癌症的免疫反应的多效性细胞因子。高水平的血清IL-6是乳腺癌患者的特征，水平升高与侵袭性疾病有关，包括更多的转移部位和对标准治疗的抵抗。IL-6基因启动子区域的遗传多态对细胞因子的转录速率具有重要的功能意义。我们建议研究这些基因多态性对结节阳性乳腺癌患者的无病生存期(DFS)和总生存期(OS)的影响。我们建议在东方合作肿瘤学小组(ECOG)进行的大型、多机构临床试验的背景下研究这一问题。这项研究是一项随机试验，在540名至少有10个阳性淋巴结的女性中，比较了标准的基于蒽环类药物的辅助化疗和大剂量化疗+自体干细胞挽救，将提供胚系DNA(来自储存的干细胞)、肿瘤标本和实质性的随访(5.7年)，以解决四个主要假设：(1)IL-6启动子基因多态性与淋巴结阳性乳腺癌患者的DFS和OS有关，(2)这种关系被肿瘤的雌激素受体状态和患者的绝经状态所改变；(3)这些多态与肿瘤受体和/或调节临床结果的信号的特定变化有关；(4)这些多态与所用治疗的毒性有关。我们将利用基于PCR的基因分型技术来鉴定IL-6启动子的以下多态性：-174G/C、-597G/A、-572G/C和-373AnTn。我们将利用免疫组织化学染色来确定肿瘤受体(IL-6R、gp130、Her2/neu、EGFR)、细胞凋亡(通过TUNEL、STAT-1、STAT-3、bCL-2)、细胞周期控制(通过MIB-1、AKT、p27)和环氧合酶-2酶活性(COX-2)的变化，所有这些都已被证明在体外与IL-6相互作用。卡普兰-迈耶和考克斯比例风险技术将被用来调查感兴趣的关联。这项研究将提供宿主环境的遗传决定因素与肿瘤行为之间相互作用的重要信息，并将提供亟需的机制数据，这些数据将对利用这些信息开发针对这种毁灭性疾病的靶向治疗方法非常重要。

DESCRIPTION (provided by applicant): Breast cancer remains a major public health problem, with over 190,000 new cases in the U.S. annually. Approximately 1/3 of these cases comprise women with disease limited to the breast and axillary lymph nodes. In these patients, aggressive multi-modality treatment can be curative in just over 50% of cases. However, current prognostic markers are unable to discriminate between those who will be cured with therapy, those who are cured without aggressive therapy and those who are destined to recur regardless of therapy. In order to improve survival and appropriately target therapies, better markers and improved understanding of the biology of recurrence is clearly needed. The proposed study seeks to expand our knowledge of breast cancer recurrence and prognosis by examining the role of host genetic characteristics on treatment outcome, and examining links between these host factors and tumor molecular characteristics. The host genetic factor of interest in this study is Interleukin-6 (IL-6), a pleiotropic cytokine involved in the host immune response to cancer. High serum levels of IL-6 are characteristic in breast cancer patients, and increased level is associated with aggressive disease, including more metastatic sites and resistance to standard therapies. Genetic polymorphisms in the promoter region of the IL-6 gene confer functional significance in transcription rates of the cytokine. We propose to examine the impact of these polymorphisms, individually and in combination, on disease-free survival (DFS) and overall survival (OS) in node-positive breast cancer patients. We propose to study this issue in the context of a large, multi-institution clinical trial conducted by the Eastern Cooperative Oncology Group (ECOG). This study, a randomized trial comparing standard anthracycline-based adjuvant chemotherapy to high dose chemotherapy with autologous stem cell rescue in 540 women with at least 10 positive lymph nodes, will provide germ-line DNA (from stored stem cells), tumor specimens and substantial follow-up (5.7 years) to address four main hypotheses: (1) that IL-6 promoter polymorphisms are associated with DFS and OS in patients with node-positive breast cancer, (2) that this relationship is modified by estrogen receptor status of the tumor and menopausal status of the patient (3) that these polymorphisms are associated with specific alterations in tumor receptors and/or signaling that mediate the clinical outcomes and (4) that these polymorphisms are related to the toxicity of the treatments administered. We will utilize PCR-based genotyping techniques to identify the following polymorphism in the IL-6 promoter: -174G/C, -597G/A, -572G/C and -373AnTn. We will utilize immunohistochemical staining to identify alterations in tumor receptors (IL-6R, gp130, Her2/neu, EGFR), apoptosis (via TUNEL, STAT-1, STAT-3, bcl-2), cell cycle control (via MIB-1, AKT, p27) and cyclooxygenase-2 enzyme activity (COX-2), all of which have been shown to interact with IL-6 in vitro. Kaplan-Meier and Cox proportional hazards techniques will be used to investigate the associations of interest. This study will provide important information on interactions between genetic determinants of the host environment and tumor behavior, and will provide much-needed mechanistic data that will be important in utilizing this information for the development of targeted therapies for this devastating disease.